c-kit Oncoprotein (CD117) Antibody Staining Protocol for Immunohistochemistry


Description: The c-kit proto-oncogene encodes a transmembrane tyrosine kinase receptor, c-kit (CD117) which is closely related to the platelet derived growth factor family. c-kit plays a role during haematopoiesis, gametogenesis and melanogenesis.


Primary Antibody

Name: c-kit Oncoprotein (CD117) Antibody

Clone: 57A5D8, Mouse anti-Human

Supplier: Novocastra Labs

Catalog Number: NCL-cKIT

Dilution: 1:200 - 1:400 using IHC-TekTM Antibody Diluent (Cat# IW-1000 or IW-1001) to reduce background and unspecific staining and serum blocking step is NOT needed.

Incubation Time/Temp: 60 minutes/room temperature

Antigen Retrieval
Device: No antigen retrieval is needed
Buffer/pH value: N/A
Heat/Cool Temperature: N/A
Heat/Cool Time: N/A

Detection Methods
Standard Method: ABC Method or LSAB Method
Enhanced Method: Polymeric Methods

Chromogen Substrate
Reagent: DAB
Incubation Time/Temperature: 1-3 minutes/room temperature

Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

Staining Pattern: Membrane/cytoplasmic
Images: Search image

Additional Information:
Tissue Type: Skin
Fixation: Acetone fixed frozen sections.
Positive Control: Normal skin, epidermal melanocytes.
Negative Control: Omit primary antibody, isotype control or absorption control
Blocking: 2-5% normal serum to reduce unspecific background staining; 0.5-3% H2O2 to block endogenous peroxidase activity; avidin/biotin to block endogenous biotin activity if necessary


Not tested on paraffin sections